Globally, approximately 170 million people are infected with the Hepatitis C Virus (HCV). HCV infection results in severe liver disease that includes chronic hepatitis, cirrhosis and hepatocellular carcinoma. Chronic HCV infection is a leading risk factor for end stage liver disease and liver transplantation. Therapy with pegylated interferon is the only available treatment yet is expensive, ineffective in many HCV patients and associated with significant side effects. Much research has been devoted to studying the replication mechanism of the HCV genome during infection. However, late post-replication events such as budding and egress have not been addressed. The recent development of cell culture systems that recapitulate the full HCV lifecycle have finally allowed viral late events to be studied. The viral-host interactions that HCV utilizes to assemble and exit from the host cell are a virtual black box. The aims of this proposal are to chtegies will be employed. Known host factors involved in the endocytic, secretory and autophagy pathways will be down regulated using RNA interference and pharmacological inhibitors to identify a role in infectious virus production. Candidate host factors implicated by such methods will be further assessed for genetic and biochemical interactions with HCV and microscopic techniques such as electron and immuno fluorescent microscopy will identify the organelles that serve as a platform for HCV assembly and release. Using biochemical and genetic approaches we will identify viral factors important for mediating late events such as assembly and egress. The Hepatitis C Virus (HCV) infects an estimated 170 million people worldwide and almost 3 million in the United States. HCV infection results in severe liver diseases such as chronic hepatitis, cirrhosis and liver cancer, and is a leading risk factor for end stage liver disease and liver transplantation. This proposal will shed light on the unknown mechanism of HCV release from infected cells, thereby creating a number of targets for potential anti-HCV drugs, which are currently limited to one form of expensive and only partially successful therapy. 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